Fig. 2: MPP+-induced SNCA transcription correlates with TRIM17 induction and ZSCAN21 stabilisation. | Cell Death & Disease

Fig. 2: MPP+-induced SNCA transcription correlates with TRIM17 induction and ZSCAN21 stabilisation.

From: ZSCAN21 mediates the pathogenic transcriptional induction of α-synuclein in cellular and animal models of Parkinson’s disease

Fig. 2

A After 6 days of differentiation, LUMHES spheroids were treated or not with 2 or 5 μM MPP+ for 24 h. Then, total RNA was extracted, and the pre-mRNA/mRNA levels of TRIM17 and the mRNA levels of TRIM41 were estimated by quantitative RT-PCR, using YWHAZ and TBP as reference genes. The graph shows mean ± SEM and individual results of five independent experiments. *: significantly different from non-treated cells (0) (one-way ANOVA followed by Dunnett’s multiple comparison test). B LUMHES spheroids were treated as in (A), and total protein extracts were analysed by western blot using antibodies against TRIM17, TRIM41 and GAPDH. The intensity of the TRIM protein bands was quantified, normalised by the intensity of the GAPDH bands and expressed relative to the values obtained with non-treated cells. The graph shows mean ± SEM and individual points from three independent experiments. *Significantly different from non-treated cells (one-way ANOVA followed by Dunnett’s multiple comparison test). C LUMHES spheroids were treated or not with 2 μM MPP+ for 24 h. Then, total protein extracts were analysed by western blot using antibodies against ZSCAN21 and GAPDH. The results were analysed as in (B). The graph shows mean ± SEM and individual points from five independent experiments. *: significantly different from non-treated cells (unpaired t-test). D LUMHES spheroids were treated as in (A). Then, total RNA was extracted, and the mRNA level of ZSCAN21 was determined and analysed as in (A). E LUMHES spheroids were treated or not with 2 μM MPP+ for 24 h and with 40 μg/ml cycloheximide (CHX) for 0 h, 16 h or 24 h, as indicated. Total proteins were analysed by western blot using antibodies against ZSCAN21 and GAPDH. For each experiment, the amount of ZSCAN21 was normalised by the level of GAPDH in each condition and plotted against CHX incubation time. Data are the mean ± SEM of four independent experiments. *Significantly different from non-treated cells at the indicated time point (two-way ANOVA followed by Sidak’s multiple comparison test).

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